Expression Of Gamma-aminobutyric Acid Research Articles

Developmental kinetics of GAD family mRNAs parallel neurogenesis in the rat spinal cord

GABA (gamma-amino butyric acid), a fast-acting synaptic transmitter in the mature CNS, is synthesized from glutamate by GAD (glutamic acid decarboxylase). We have developed an ultrasensitive PCR technique to quantify the expression of GAD-related mRNAs during the development of the rat cervical spinal cord and have localized them using in situ hybridization. GAD65, GAD67, and an alternatively spliced variant of GAD67, EP10, were quantified each day from embryonic (E) day 11 through E21, and at postnatal days 0, 7, 14, and adult. GAD65 and GAD67 mRNAs were detected at E11 and increased exponentially over three orders of magnitude during embryonic development, then declined approximately threefold in the first-2 postnatal weeks. While the exponential growth phase coincided with the progressive appearance of GAD67 in situ signals in both the ventral and dorsal cord, the postnatal decline coincided with the virtual disappearance of expression in the ventral region. EP10 expression was prominent in the embryo, then declined markedly together with the mRNA encoding the neuroepithelial stem cell marker, nestin. The concerted appearance of GAD-related mRNAs paralleled transcripts encoding neuronal markers (light and heavy neurofilaments) and also closely correlated with the expression of GABA, mRNAs encoding GABAA receptor subunits, and depolarizing responses to GABA. We have used the results on GAD-related mRNA expressions to formulate a simple, minimal mathematical model that accounts for their kinetics in terms of positive and negative feedback loops.

Trimethyltin-induced expression of GABA and vimentin immunoreactivities in astrocytes of the rat brain.

Adult Sprague-Dawley rats were given a single dose of trimethyltin chloride (TMT). Three days following treatment, a neuronal alteration was observed in the CA3c pyramidal cell layer of hippocampus which was not accompanied by any apparent astrocyte reaction. At 1 as well as 2 weeks after treatment, a gliosis in hippocampus, piriform, and entorhinal cortices was detected by glial fibrillary acidic protein (GFAP) immunohistochemistry. Concomitant with the enhanced astrocytic GFAP, astrocytes were swollen and expressed immunoreactivity to vimentin and gamma-aminobutyric acid (GABA). The astrocytic GABA immunoreactivity may reflect a trimethyltin-induced alteration in astrocyte phenotype, or alterations in compartmentalization and/or metabolism of GABA.

The development of GABA immunoreactivity in the retina of the zebrafish (Brachydanio rerio).

The goal of this study was to determine the pattern of gamma-aminobutyric acid (GABA) expression in the retina and optic nerve of the zebrafish (Brachydanio rerio) during embryonic development. Zebrafish embryos were fixed at intervals between 1 and 4 days postfertilization, and semithin plastic sections were prepared for postembedding immunocytochemistry with antisera against GABA. Sections were also prepared from several adult zebrafish eyes for comparison. GABA immunoreactivity first appeared in the optic nerve at 2 days postfertilization, and by 2.5 days the inner nuclear layer (INL), inner plexiform layer (IPL), retinal ganglion cell layer, and optic nerve were all positive for GABA. The GABA expression in the retinal ganglion cell layer and optic nerve was transient, however, and these structures were largely unlabeled by 4 days postfertilization. The pattern of GABA immunoreactivity at 4 days resembled that seen in the adult zebrafish: A large population of presumptive amacrine cells was labeled at the base of the INL, and the IPL was positive for GABA, as were occasional cells in the ganglion cell layer. Horizontal cells, particularly at the retinal margins, were also GABA positive beginning at about 3 days postfertilization. The transient expression of GABA in retinal ganglion cells and their axons during the period when synaptic contacts are being established both within the retina and between the retina and central targets suggests that GABA may have a role in the development of this system, in addition to serving as a classical neurotransmitter.

Interaction between enkephalin and gamma-aminobutyric acid in the chicken retina: a double-label immunoelectron microscopic analysis.

In the present study, double-label immunoelectron microscopy was used to examine the synaptic relationships between amacrine cell populations in the chicken retina that contain either enkephalin or gamma-aminobutyric acid (GABA) or both enkephalin and GABA. The objectives of the present study were twofold. First, the ultrastructural features and synaptic organization of enkephalin and enkephalin/GABA amacrine cells were compared. Second, the synaptic interactions between these populations and the population of GABA amacrine cells were examined. A total of 475 synaptic arrangements were observed to involved enkephalin or enkephalin/GABA amacrine cell processes. The synaptic relationships of enkephalin and enkephalin/GABA amacrine cells were quite similar. Each population was pre- and postsynaptic to amacrine cells, postsynaptic to bipolar cells, and presynaptic to processes possibly originating from ganglion cells. A substantial percentage of each population's pre- and postsynaptic relationships were with the processes of GABAergic amacrine cells. Moreover, when enkephalin and enkephalin/GABA amacrine cell processes were postsynaptic to bipolar cells, their dyadic partner was observed frequently to be a GABA amacrine cell process. The present study suggests a diversity in the population of chicken enkephalin amacrine cells with respect to their expression of the classical inhibitory transmitter GABA. Moreover, a functional relationship between enkephalinergic and GABAergic pathways is indicated by studies showing that both enkephalin and enkephalin/GABA amacrine cells exhibit substantial synaptic interaction with GABA amacrine cells.

The GABAergic nervous system of Caenorhabditis elegans.

gamma-Aminobutyric acid (GABA) is the most abundant inhibitory neurotransmitter in vertebrates and invertebrates. GABA receptors are the target of anxiolytic, antiepileptic and antispasmodic drugs, as well as of commonly used insecticides. How does a specific neurotransmitter such as GABA control animal behaviour? To answer this question, we identified all neurons that react with antisera raised against the neurotransmitter GABA in the nervous system of the nematode Caenorhabditis elegans. We determined the in vivo functions of 25 of the 26 GABAergic neurons by killing these cells with a laser microbeam in living animals and by characterizing a mutant defective in GABA expression. On the basis of the ultrastructurally defined connectivity of the C. elegans nervous system, we deduced how these GABAergic neurons act to control the body and enteric muscles necessary for different behaviours. Our findings provide evidence that GABA functions as an excitatory as well as an inhibitory neurotransmitter.

Genes required for GABA function in Caenorhabditis elegans.

gamma-Aminobutyric acid (GABA) neurotransmission is widespread in vertebrate and invertebrate nervous systems. Here we use a genetic approach to identify molecules specific to GABA function. On the basis of the known in vivo roles of GABAergic neurons in controlling behaviour of the nematode Caenorhabditis elegans, we identified mutants defective in GABA-mediated behaviours. Five genes are necessary either for GABAergic neuronal differentiation or for pre- or postsynaptic GABAergic function. The gene unc-30 is required for the differentiation of a specific type of GABAergic neuron, the type-D inhibitory motor neuron. The gene unc-25 is necessary for GABA expression and probably encodes the GABA biosynthetic enzyme glutamic acid decarboxylase. The genes unc-46 and unc-47 seem to be required for normal GABA release. Finally, the gene unc-49 is apparently necessary postsynaptically for the inhibitory effect of GABA on the body muscles and might encode a protein needed for the function of a GABAA-like receptor. Some of these genes are likely to encode previously unidentified proteins required for GABA function.

Differential expression of GABAA/benzodiazepine receptor beta 1, beta 2, and beta 3 subunit mRNAs in the developing mouse cerebellum.

Gamma aminobutyric acid (GABA) is the major inhibitory neurotransmitter in the mammalian cerebellum. Cerebellar granule, Purkinje, and deep nuclear neurons are known to receive GABAergic afferents. Since GABA exerts its inhibitory effects via GABA receptors, it is of interest to determine the temporal relationship between the formation of GABAergic synapses and the expression of genes coding for the GABA receptor. In a previous study, we have examined the developmental expression of binding sites for [3H]muscimol, which binds with high affinity to the beta subunits of the GABAA/benzodiazepine (GABAA/BZ) receptor. In the present study, [35S]cRNA probes were used to examine the appearance and distribution of GABAA/BZ beta 1, beta 2, and beta 3 subunit mRNAs in the developing C57BL/6 mouse cerebellum by in situ hybridization. In the adult cerebellum, the distribution of the three subunit mRNAs was clearly different, despite considerable overlap, and their temporal expression differed throughout postnatal development. The beta 1 hybridization signal appeared within the cerebellar cortex during the second postnatal week as a discrete band at the interface of the molecular and granule cell layers. Grains were distributed diffusely over small densely staining cells surrounding the Purkinje cells; relatively few grains were visible over Purkinje cell bodies themselves. This distribution may reflect an association with Bergmann glia or basket cells. The beta 2 and beta 3 hybridization signals were present considerably earlier than that of the beta 1 mRNA. The beta 2 signal was present at birth in the molecular/Purkinje cell layer; as development progressed, the signal became increasingly intense over both granule and Purkinje cells. At birth, the beta 3 subunit mRNA was present in the external germinal and molecular layers, later becoming largely localized within the granule cell layer. Dense beta 2 and beta 3 cRNA probe labeling was present over the adult granule cell layer. Moderate levels of beta 2 signal were seen over Purkinje cell bodies; considerably less labeling was observed with the beta 3 probe. The adult distribution of beta 2 and beta 3 cRNA probes showed good spatial correspondence with the known GABAA receptor beta subunit markers, [3H]-muscimol and the mAb 62-3G1 antibody, each being present within the granule cell layer. Our results indicate that the temporal expression of GABAA/BZ receptor beta subunit messages within a given cell type may be independently regulated, and that acquisition of the beta 2 and beta 3 mRNAs occurs before these cells become integrated into mature synaptic circuits.

Injury-induced reorganization of somatosensory cortex is accompanied by reductions in GABA staining.

When a portion of primary somatosensory cortex is deprived of its normal inputs by peripheral nerve transection, intact skin surfaces represented in surrounding cortex come to activate the deprived zone within 2 months. We found that this cortical reorganization was accompanied by a marked decrease in the antibody staining of gamma-aminobutyric acid (GABA) within the deprived sector of cortex in monkeys surviving nerve injury for 2-5 months. In contrast, there were no apparent changes in cytochrome oxidase reactivity in the deprived cortex of these same monkeys. Reduced levels of inhibition could allow previously unexpressed connections to become potent. Thus, the regulation of the expression of GABA appears to be one mechanism for maintaining and altering cortical representations.

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GABA (gamma-amino butyric acid), a fast-acting synaptic transmitter in the mature CNS, is synthesized from glutamate by GAD (glutamic acid decarboxylase). We have developed an ultrasensitive PCR technique to quantify the expression of GAD-related mRNAs during the development of the rat cervical spinal cord and have localized them using in situ hybridization. GAD65, GAD67, and an alternatively spliced variant of GAD67, EP10, were quantified each day from embryonic (E) day 11 through E21, and at postnatal days 0, 7, 14, and adult. GAD65 and GAD67 mRNAs were detected at E11 and increased exponentially over three orders of magnitude during embryonic development, then declined approximately threefold in the first-2 postnatal weeks. While the exponential growth phase coincided with the progressive appearance of GAD67 in situ signals in both the ventral and dorsal cord, the postnatal decline coincided with the virtual disappearance of expression in the ventral region. EP10 expression was prominent in the embryo, then declined markedly together with the mRNA encoding the neuroepithelial stem cell marker, nestin. The concerted appearance of GAD-related mRNAs paralleled transcripts encoding neuronal markers (light and heavy neurofilaments) and also closely correlated with the expression of GABA, mRNAs encoding GABAA receptor subunits, and depolarizing responses to GABA. We have used the results on GAD-related mRNA expressions to formulate a simple, minimal mathematical model that accounts for their kinetics in terms of positive and negative feedback loops.